Polyunsaturated linear aldehydes and their derivatives with anti-radical and anti-tumoral activity

ABSTRACT

The invention relates to polyunsaturated linear aldehydes and derivatives thereof which can be extracted from the feathers or tissues of parrots of prepared synthetically with antioxidant, antitumoral and anti-inflammatory activity, and which are useful in the prevention and treatment of cardiovascular, inflammatory, atherosclerotic, proliferative cell and tumour damage and the prevention of alterations caused by ageing.

This invention relates to polyunsaturated linear aldehydes and theirderivatives with anti-radical and anti-tumoral activity.

More particularly, this invention relates to the use of polyunsaturatedlinear aldehydes which can be extracted from parrot feathers and tissuesor prepared synthetically, and of derivatives of said aldehydes such asfatty esters of alcohols obtained by reduction of the aldehyde group, orderivatives of inclusion of said aldehydes in cyclodextrins, asantioxidant, antitumoral and anti-inflammatory agents.

This invention also relates to novel derivatives of said aldehydes, suchas some alcohols obtained by reduction of the aldehyde group, fattyesters of alcohols and derivatives of inclusion of aldehydes incyclodextrins. Said polyunsaturated aldehydes and the derivativesthereof will hereafter be generically referred to as “parrodienes”.

It has been found that the parrodienes of the invention possessantioxidant, anti-tumoral and anti-inflammatory activity and aretherefore useful in preventing the damage caused by free radicals, andin particular in the prevention and treatment of cardiovascular,inflammatory, atherosclerotic, proliferative cell and tumour damages andthe prevention of alterations caused by ageing.

Numerous natural products of plant origin, present in the diets ofvarious populations, possess a preventive or curative activity againstnumerous organic disorders and alterations characteristic of variousdiseases.

These therapeutic properties are generally indicated by folk tradition,and their validity has been investigated by modern chemical, biological,pharmacological and clinical techniques.

Far fewer natural products of animal origin included in the diet havebeen found to have therapeutic or preventive properties againstparticular disorders or diseases.

Known examples are polyunsaturated fatty acids extracted from fish (1,2, 3), glycosaminoglycans extracted from the cartilage of animals suchas the shark (3, 4, 5, 6, 8), glycoproteins such as lactoferrinextracted from milk or colostrum (9, 10), and lipid extracts of molluscssuch as oysters (11, 12).

However, the biological activity of the polyunsaturated linear aldehydespresent in the feathers and tissues of parrots is wholly unexplored.

Folk traditions of Brazilian and Venezuelan people hand down thatexternal application of the feathers of these birds heals skininfections, burns and poisonous insect bites, and that eating their meatcures intestinal disorders, infections and tumours. It is known that thecoloured plumage of birds have numerous types of carotenoids which areabsorbed through the diet and can bond directly with the keratin in thefeathers or undergo a complex metabolic transformation.

Lutein, zeaxanthin and beta-cryptoxanthins are carotenoids frequentlyfound in foods (berries, fruit, seeds, flowers and insects), which canbe absorbed by tissues and feathers with no metabolic modifications totheir structure, whereas other compounds, such as picofulvins, are theresult of molecular modification (13, 14, 15, 16). The psittacofulvinsor the mixture of parrodienes (also indicated here by the name ofParrodin) are the result of metabolic changes typical of various speciesof parrot (Poicephalus rufiventris, Ara macao, Ara manilata, Araararauna, Psittacus erithacus, Aratinga canicularis, Aratingaacuticaudata, Psittacula krameri, etc.).

The parrodienes according to the invention have an antioxidant andanti-tumoral activity and in general a protective effect against thefree radicals, which may explain the exceptional longevity of theseanimals (they can live for up to 100 years), their learning ability, andabove all their lack of diseases, especially tumours.

The compounds of the invention, with the general formula (I)

wherein

-   -   n=2-7,    -   R=CHO, CH₂OH, CH₂O—CO—R′, wherein —CO—R′ is the residue of a        fatty acid with 12-22 carbon atoms,

were first obtained (R=CHO) as polyene aldehydes, the synthesis of whichwas described for the first time by Kuhn in 1937 (17, 18). Aldehydeswith an odd number of conjugated double bonds were obtained byautocondensation of crotonaldehyde in the presence of the catalystpiperidinium acetate (scheme 1A). Aldehydes with an even number ofdouble bonds were obtained by condensation between crotonaldehyde andacetaldehyde, again in the presence of the catalyst piperidinium acetate(scheme 1B).

The corresponding alcohols (R=CH₂OH) were obtained by reduction of thealdehydes with NaBH₄; the esters were obtained by esterification withthe required acyl halides, especially chlorides.

Examples of preparation of the compounds according to the invention areset out below.

EXAMPLE 1A Preparation of Aldehydes with an Odd Number of Double Bonds

Formula (I), n=3: 2,4,6-octatrienal

 n=5: 2,4,6,8,10-dodecapentaenal

 n=7: 2,4,6,8,10,12,14-hexadecaheptaenal

250 ml of crotonaldehyde are added to a 1-litre flask and keep undermagnetic stirring for 15 minutes under nitrogen flow. 2.5 ml ofpiperidine and 2.5 ml of acetic acid are slowly dropped therein (thereaction is exothermic) and the mixture is left under magnetic stirringin a nitrogen atmosphere at 50° C. for 30 minutes.

The reaction mixture is cooled in an ice bath and added with 600 ml ofethyl ether, still under magnetic stirring. The brown precipitate formedis filtered and recrystallized from toluene: 120 mg of2,4,6,8,10,12,14-hexadecaheptaenal is obtained.

The red ether solution is extracted with 200×5 ml of distilled water towash away the unreacted surplus crotonaldehyde; the organic phase isdried over sodium sulphate and evaporated under pressure (30 mmHg). Thebrownish-red residue is taken up with 30 ml of 80% methanol and left inthe refrigerator overnight at 3° C. 2,4,6,8,10-dodecapentaenal is thusseparated by precipitation, and is recrystallized from isopropanol (2.5g).

The methanol solution is evaporated and distilled at low pressure (3mmHg): the 2,4,6-octatrienal separates at 55°-60° C. and is purified ona silica gel flash chromatography column, eluting with a mixture ofpetroleum ether/ethyl acetate (ratio 98:2). 5 g of 2,4,6-octatrienal isobtained.

TLC: Hexane 7/Acetone 3

EXAMPLE 1B Preparation of Aldehydes with an Even Number of Double Bonds

Formula (I), n = 2: 2,4-hexadienal n = 4: 2,4,6,8-decatetraenal n = 6:2,4,6,8,10,12-tetradecahexaenal

190 ml of acetaldehyde and 140 ml of crotonaldehyde are placed in a1-litre flask, and kept under magnetic stirring for 30 minutes undernitrogen flow. 2 ml of piperidine and 1.4 ml of acetic acid are slowlydropped therein, and the mixture is left under magnetic stirring in anitrogen atmosphere for 18 hours. 500 ml of ethyl ether are added to thered solution; the precipitate is filtered, then recrystallized fromtoluene. The precipitate obtained is 2,4,6,8,10,12,14-hexadecaheptaenal(140 mg), whereas 2,4,6,8,10,12-tetradecahexaenal (25 mg) is isolatedfrom the evaporated toluene phase.

The red ether solution is washed with 200×5 ml of distilled water, driedover sodium sulphate and left to stand at −20° C. for 12 hours.2,4,6,8,10,12-tetradecahexaenal thus separated by precipitation isfiltered and washed with ether (50 mg).

The residue obtained from evaporation of the organic phase is subjectedto fractional distillation: unreacted crotonaldehyde is obtained at 21°C. (30 mmHg), 2,4-hexadienal at 26° C. (3 mmHg, 5.2 g) and2,4,6-octatrienal at 55-60° C. (3 mmHg). The residue is taken up with 30ml of 80% methanol; 2,4,6,8-decatetraenal is separated by precipitationat −20° C. and then filtered and recrystallized from hexane (420 mg).

The fraction of distillate containing 2,4,6-octatrienal is purified on asilica gel flash chromatography column, eluting with a mixture ofpetroleum ether/ethyl acetate (ratio 98:2), to obtain 3 g of2,4,6-octatrienal.

TLC: Hexane 7/Acetone 3.

EXAMPLE 2 Preparation of 2,4,6,8,10,12-tetradecahexaenol

2.2 mmols of 2,4,6,8,10,12-tetradecahexaenal are dissolved in 20 ml ofanhydrous ethanol in a flask under nitrogen flow; 3.3 mmols of NaBH₄,solubilized in 5 ml of anhydrous ethanol, are dropped into the solution.The mixture is left at room temperature under magnetic stirring in anitrogen atmosphere, checking periodically with TLC (ethyl acetate).After reacting for 5 hours, 30 ml of distilled water are added and themixture is extracted with diethyl ether; the combined organic phases aredried over anhydrous sodium sulphate and evaporated under pressure (30mmHg). The resulting crude product is purified by crystallisation fromethyl acetate. 2,4,6,8,10,12-tetradecahexaenol is obtained with a 60%yield.

The alcohols described in the table below were prepared in accordancewith the procedure described above. Reaction % No. NAME timePurification yield 2 2,4-hexadienol⁽¹⁹⁾ 1 hour 84 32,4,6,-octatrienol⁽¹⁹⁾ 1½ hours Recrystallized from 70 petroleum ether 42,4,6,8-decatetraenol⁽¹⁹⁾ 2 hours Recrystallized from ethanol 70 52,4,6,8,10-dodecapentaenol^((20,21)) 3 hours Recrystallized from ethyl78 acetate 6 2,4,6,8,10,12,14-hexadecaheptaenol 5 hours Chromatographycolumn, 40 eluting with ethyl acetate

EXAMPLE 3 Preparation of 2,4,6-octatrienyl palmitate

2 mmols of 2,4,6-octatrienol under nitrogen stream are solubilized inanhydrous CHCl₃ not stabilized with ethanol; 2 mmols of anhydroustriethylamine are dropped into the solution. Keeping the flask undermagnetic stirring in an ice bath, 3 mmols of palmitoyl chloride areslowly dropped into 10 ml of CHCl₃. When the addition is finished, thereaction mixture is brought to room temperature, checking periodicallywith TLC (ethyl acetate). The reaction is complete after approx. 5hours. The reaction solution is extracted with distilled water and thenwith a saturated aqueous solution of NaHCO₃ dried over anhydrous sodiumsulphate and evaporated (30 mmHg).

2,4,6-Octatrienyl palmitate with the following spectroscopiccharacteristics is obtained (yield 91%):

IR (KBr): 2920, 2850, 1740, 1560, 1264, 996.

¹H-NMR (CDCl₃): ppm 6.372-6.040 (4H, m, H₃₋₆); 5.791-5.959 (2H, m, H₂,H₇); 4.634 (2H, d, J_(1.2)=5.64, CH₂O); 2.326 (2H, t, J_(5.6)=7.3,CH₂CO); 1.816 (3H, d, J_(8,7)=7.3, CH₃—CH═); 1.430-1.157 (26H,m, CH₂);0.928 (3H, t, J=6.23, CH₃).

¹³C (CDCl₃): ppm 173.648 COO; 134.687 C₄; 134.454 C₃; 134.454 C₂;134.454 C₅; 134.454 C₆; 134.454 C₇; 64,709 CH₂O; 34.430-22.738 CH₂;18.309 CH₃—CH═; 14.134 CH₃.

The esters described in the table below were prepared in accordance withthe procedure described above. Reaction % No. NAME time yield 22,4-hexadienyl palmitate  4 hours 94 3 2,4,6,8-decatetraenyl palmitate24 hours 86 4 2,4,6,8,10-dodecapentaenyl palmitate 24 hours 84 52,4,6,8,10,12-tetradecahexaenyl palmitate 24 hours 60 62,4,6,8,10,12,14-hexadecaheptaenyl palmitate 24 hours 40

EXAMPLE 4 Preparation of Inclusions of Aldehydes in Cyclodextrins

Equimolar quantities of polyene aldehyde and α or β-cyclodextrin aremixed and worked in a mortar until a homogenous mixture is obtained. Themixture is worked for 30 minutes, added with 2 ml of distilled water intwo successive times; the semi-liquid mixture thus obtained is placedinto a flask kept in a nitrogen atmosphere overnight, and resuspend in200 ml of warm distilled water (40° C.). The coloured suspension is keptunder magnetic stirring at 40° C. for 20 minutes, then filtered whilehot under vacuum (30 mmHg) through a sintered glass filter. A smallamount of the yellow solution, stored at 4° C., does not contain anysediment or precipitate, even after several days. The product to becharacterized is obtained by evaporation under vacuum (30 mmHg) of theaqueous solution.

Inclusions of polyene aldehydes containing 3, 4, 5, 6 and 7 conjugateddouble bonds in α and β-cyclodextrin were prepared in accordance withthis procedure. Each complex obtained was characterized by IR, Raman andUV/vis spectroscopy.

2,4,6-octatrienal included in α-cyclodextrin

IR (cm⁻¹) 2929; 1680; 1641; 1413; 1384; 1298, 1244,1155; 1078; 1030;951.

Raman (cm⁻¹) 1633, 1611; 1160; 1129.

UV/vis in H₂O: λ(nm): 277.5

 ε(1 cm⁻¹ mol⁻¹):705

2,4,6-octatrienal included in β-cyclodextrin

IR(cm⁻¹) 2927; 1667; 1644; 1417; 1369; 1302; 1247; 1158; 1080; 1028;947.

Raman (cm⁻¹) 1678; 1636; 1126.

UV/vis in H₂O: λ(nm): 276.5

 ε(1 cm⁻¹ mol⁻¹): 604

Biochemical and pharmacological experiments conducted on the compoundsaccording to the invention have led to a characterization whichindicates that they have a potential role in the prevention andtreatment of various common disorders.

Their activity against the lipoperoxidation induced by CCl₄ on isolatedrat hepatocytes, and against the oxidative phenomena induced by H₂O₂ onthe phaeochromocytoma cell (PG 12), has been demonstrated by in vivo andin vitro tests.

Their ability to prevent hydroperoxide damage has also been demonstratedin a suspension of red blood cells placed in contact with hydrogenperoxide.

This test demonstrated that the compounds according to the inventionhave an elective inhibitory capacity against the damage caused by thetoxic action of hydroperoxides.

Inhibition of collagen-induced platelet aggregation, and even moresignificantly, a reduction in reperfusion damage in the heart of therat, has been found in the experimental cardiovascular field.

Lastly, the antiproliferative activity of the compounds according to theinvention has been demonstrated on type MCF 7 and SHSY-SY tumour cellsand on type DHL4 and HL60 cells.

Protection Against H₂O₂-Induced Oxidation

A culture of phaeochromocytoma cells (PC-12) containing 3×10⁻⁵ Mcells/ml was used; according to the method described by Nordman (NordmanR., Free Rad. Biol. Med., 1227, 1996) it was subjected to H₂O₂ at theconcentration of 0.1 mM for 30 min. 100 or 200 mcg/cc of “parrodin” (ie.the mixture of aldehydes with formula (I), wherein R=CHO) was added tothe cell culture at the start of the experiment.

After 24 hours' incubation it was observed that with the peroxidationinduced by H₂O₂ the survival rate was 30%, whereas the survival rate ofthe cells incubated with parrodin as well as H₂O₂ was 50% at theconcentration of 100 mcg/cc and 90% at the concentration of 200 mcg/cc,thus demonstrating that parrodin significantly protects against theperoxidative damage caused by H₂O₂.

Tests of Lipoperoxidation Induced by CCl₄.

The lipoperoxidative and toxic effect induced by CCl₄ (100 mg/L⁻¹) wasevaluated on rat hepatocytes isolated according to the techniquedescribed by Segler (Segler P. O., Methods Cell. Biol. Chem., 264, 4747,1989). CCl₄ is known to cause lipoperoxidation of the cell membraneswhich can lead to cell necrosis (Slater T. F., Philos Trans. R. Soc.Lond. (Biol) 311, 633, 1985; Berger M. L., Hepatology, 6, 36, 1996;Tribble D. L., Hepatology, 7, 377, 1987).

The damage to the cell membrane caused by CCl₄ and the protective effectperformed were measured by assaying alanine aminotransferase (AlaAT) andaspartate aminotransferase (AspoAT) on the supernatant liquid of thecell culture (Auto-biochemistry Assay System-Beeckman 700-Encore-2).

Cytological tests on the hepatocytes were performed under the optical orelectronic microscope after they had been fixed in formalin orglutaraldehyde. The results of these tests demonstrated that theincrease in AlaAT and AspAT concentrations caused by CCl₄ is reduced bythe presence of parrodin.

The histological tests also demonstrated the protection provided byparrodin. The cell membranes and nucleus of the treated hepatocytes,unlike the controls, appeared almost intact, and the mitochondria andthe number of ribosomes also appeared normal.

Test of Lipoperoxidation Induced by Hydrogen Peroxide on Red Blood Cells

Red blood cells were extracted from the venous blood of healthyvolunteers; after centrifugation and washing in saline buffered with PBSphosphates (0.15M, pH 7.4) they were diluted with 10 cc of a solutioncontaining 10⁻³M of PBS-azide, and the haemoglobin concentration wasmeasured with Drabkin's reagent. Lipid peroxidation was induced byexposing a cell expansion contained in 5 cc of PBS-azide with a finalhaemoglobin concentration of 3.75 mg/ml to hydrogen peroxide (20 mM ofhydrogen peroxide per ampoule containing 5 cc of cell suspension) andincubating them at 37° for one hour.

Peroxidation was determined after one hour according to the Stocks andDormandy method (Stocks J., Dormandy T. L., Brit. J. Haematology, 29,95, 1971) that measures the formation of malonaldehyde which, incombination with thiobarbituric acid (TBA), forms a coloured chromogenwith absorbance at 532 nm (Bird R. P., Methods Enzymol, 105, 299, 1984).

The antilipoperoxidative activity of parrodin was evaluated byintroducing it into tubes containing the erythrocyte suspension at thedose of 100 mcg/ml.

The MDA measurement after one hour's incubation with hydrogen peroxidedemonstrates a highly significant reduction of MDA formation byparrodin, and consequently evident protection against lipoperoxidationdamage.

Tests on Hepatoma Cell Growth

Cells originating from human hepatoma Hep. 3B (ATCC—Manassas, U.S.A.)were used, and the proliferation processes were observed on these cellsafter they were placed in contact with parrodin. The hepatoma cells wereplaced on a suitable culture medium (Gilico Laboratories, Santa Clara,Calif., U.S.A.) added with 10% of inactivated bovine serum in thepresence of benzylpenicillin and streptomycin.

The cells (8×107 cells/l) were kept in the culture medium at 37° for 24hours. After this period, the culture medium was replaced with a culturemedium containing parrodin (5 mg or 10 mg/cc). The time needed to doublethe initial number of cells was evaluated with the control cells andthose placed in contact with parrodin. It was found that the time neededto double the number of control cells was approx. 20 hours, while thetime needed to double the number of cells placed in contact withparrodin was 28 or 36 hours, depending on the concentration used.

The antiproliferative effect performed by parrodin was thusdemonstrated.

Tests of Antiproliferative Activity

Antiproliferative activity was evaluated with teleocidin, a compoundwhich, like the phorbol myristates, causes keratosic tumoral processesin the skin (Fujiki H., Biochem. Biophys. Res. Comm., 90, 976, 1979),with an increase in the ornithine decarboxylase enzyme proportional tothe severity of the lesion induced.

Teleocidin was administered subcutaneously on the shaved back of themouse at the dose of 5 mcg/mouse, dissolved in 0.2 cc of aqueoussolution. Parrodin was administered subcutaneously to those animals forthree days prior to the test, at the dose of 10 mg or 20 mg/mouse.Alternatively, 0.3 ml of a dispersion of parrodin in lanolin, equal to aconcentration of 50 mg or 100 mg/ml, was applied to the shaved skin ofthe mouse, and the treated area was protected by an occlusive dressing.

Ornithine decarboxylase was assayed on the homogenised epidermis of thetreated animals five hours after the teleocidin injection following theO'Brien and Nakardate method (O'Brien T. G., Cancer Res., 35, 1662,1975—Nakardate T., 42, 2841, 1982).

The protein concentration of the epidermal extract was measuredaccording to the Lowry method (Lowry O. H., J. Biol. Chem, 193, 265,1951).

Ornithine decarboxylase activity was measured in nMol of CO₂/60 min./mgof protein.

These tests demonstrate that the oral and topical cutaneousadministration of parrodin reduces the ornithine decarboxylase activityof the skin to a highly significant extent, up to 50% more than thecontrols, in addition to the keratosic reaction.

Platelet Aggregation Tests

Platelet aggregation was determined on platelet-rich plasma (PRP); thenumber of platelets was counted with a CH58 oH platelet counter (Delcan)and made up to 300,000 platelets/ml with platelet-poor plasma (PPP).

Platelet aggregation was induced by adding collagen (2.5 ng/ml) andevaluated photometrically with an aggregometer according to thetechnique described by Born (Born G. V. R., Nature 194, 927, 1962).

After 10 minutes' incubation with Parrodin (2.5 ng/ml and 5 mg/ml), 55%and 75% inhibition of platelet aggregation respectively was obtained.

UV-Induced Erythema Prevention Tests

UV-induced erythema was caused by applying a UV lamp (Hanoivna Kramager)to the ear of a guinea pig for 30 sec; the lamp transmitted rays with awavelength of 200 to 400 mm, which consequently included UV-B, UV-C andUV-A rays, through a special filter.

Excipients only or a suspension containing 5 or 10%2,4,6,8,10,12,14-hexadecaheptaenol palmitate were spread on the ear ofthe test animals, and the temperature increase resulting from theUV-induced erythema was measured with a thermoelectric thermometer afterapprox. 3 hours in the control animals and the test animals. It wasfound that the 10% suspension inhibited the appearance of heat anderythema.

The set of tests performed indicates that the polyunsaturated linearcompounds according to the invention possess the biologicalcharacteristics common to carotenoids of plant or other origin, anddemonstrate a particular protective activity against hydroperoxides andfree radicals.

They consequently seem likely to be particularly useful in theprevention and treatment of all the organic changes caused by theactivation of free radical production. They can be used as dietsupplements, medicaments or cosmetics in the prevention or treatment ofalterations of cardiovascular origin, such as heart attack, stroke andatherosclerosis, or in the prevention and treatment of tumours and thevarious disorders associated with tissue aging. In cosmetology inparticular, they can be used to treat lesions caused by ultraviolet raysand skin aging processes, inflammatory or degenerative reactions.

Depending on their use, the compounds according to the invention can beemployed alone or in association with one another, or in associationwith other carotenoids or compounds with a similar, complementaryactivity, such as organic or inorganic antioxidants, vitamins,aminoacids, enzymes or other products with nutritional characteristicsor characteristics available in cosmetology.

The compounds according to the invention may be formulated in the formof tablets, granulates for oral use or ampoules for parenteral use, orin the form of ointments, creams or lotions for topical cosmeticcutaneous use.

The formulations will be prepared according to conventional techniques,using excipients suitable for pharmaceutical or cosmetological use.

Some examples of formulations according to the invention are set outbelow.

Formulation Example 1 Formulations for oral use (Tablets, capsulesetc.) - Composition of each one: “Parrodin” 10 mg “Parrodin”cyclodextrin 20 mg with excipients and additives normally acceptable inpharmaceutical technology.

Formulation Example 2 For parenteral use (ampoules, vials) - content/ccCyclodextrin - dodecapentaenal 5 mg hexadecapentaenal complex

Formulation Example 3

For topical use

(cream, ointment, lotions)—content/cc

Parrodienes

5% cyclodextrin—

dodecapentaenal hexadecapentaenal complex

Formulation Example 4 Complexes based on Parrodienes for oral use“Parrodin” cyclodextrin complex  5 mg Lycopene  5 mg β-carotene  1 mgVit. E  5 mg Vit. C 50 mg Coenzyme Q₁₀ 20 mg Selenium 50 mcg “Parrodin”cyclodextrin complex  5 mg Alpha-lipoic acid 50 mg Coenzyme Q₁₀ 30 mgVit. C 50 mg Vit. E  5 mg Vit. B₁  5 mg

Formulation Example 5 Complex based on Parrodienes for cosmetic useCream containing the following in each cc “Parrodin” cyclodextrincomplex  5 mg Melatonin 10 mg Alpha-lipoic acid 50 mg Vit. E 10 mg

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1. Use of the compounds with the general formula (I):

wherein n=2-7, R=CHO, CH₂OH, CH₂O—C—(O)R′, wherein —CO—R′ is the residueof a fatty acid with 12-22 carbon atoms, alone or in a mixture thereof,for the preparation of medicaments for the prevention and treatment ofdisorders caused by activation of free radical production.
 2. Use asclaimed in claim 1, wherein the disorders induced by activation of freeradical production are alterations of cardiovascular origin,atherosclerosis, disorders associated with tissue ageing or tumours. 3.Use as claimed in claim 2, wherein the alterations of cardiovascularorigin are heart attack and stroke.
 4. Use of the compounds as claimedin claim 1 to prepare cosmetic formulations for the treatment of damagecaused by ultraviolet rays, skin ageing processes and inflammatory ordegenerative reactions.
 5. Use of the compounds as claimed in claim 1 toprepare diet supplements.
 6. As a novel compound,2,4,6,8,10,12-tetradecahexaenol of formula (I), wherein n=6 and R=CH₂OH.7. As a novel compound, 2,4,6,8,10,12,14-hexadecaheptaenol of formula(I), wherein n=7 and R=CH₂OH.
 8. As novel compounds, the compounds ofgeneral formula (I), wherein n=2, 3, 4, 5 or 6 and R=CH₂O—CO—R′, wherein—CO—R′ is the residue of palmitic acid.
 9. Inclusion compounds ofcompounds with formula (I) in cyclodextrins

wherein n=2-7 and R=CHO.
 10. Pharmaceutical compositions containing apharmacologically or cosmetically effective amount of at least onecompound as claimed in claim
 1. 11. Pharmaceutical compositions asclaimed in claim 10, further containing other compounds with a similaror complementary activity, selected from the group consisting of organicor inorganic antioxidants, vitamins, aminoacids and enzymes. 12.Pharmaceutical compositions as claimed in claim 10 in the form oftablets or granulates for oral use, ampoules for parenteral use, orointments, creams and lotions for topical cosmetic cutaneous use. 13.Pharmaceutical compositions containing a pharmacologically orcosmetically effective amount of at least one compound as claimed inclaim
 6. 14. Pharmaceutical compositions as claimed in claim 11 in theform of tablets or granulates for oral use, ampoules for parenteral use,or ointments, creams and lotions for topical cosmetic cutaneous use.